AFFINITY ISOLATION & I-DIRT MS ANALYSIS OF E COLI O157:H7 SAKAI RNA POL COMPLEX

这个子项目是众多研究子项目之一

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Bacteria contain a single multi-subunit RNA polymerase that is responsible for the synthesis of all RNA. Previous studies on the Escherichia coli K-12 laboratory strain had identified a group of effector proteins that interact directly with RNA polymerase to modulate the efficiency of transcription initiation, elongation or termination. Here we have used a rapid affinity isolation technique to isolate RNA polymerase from the pathogenic Escherichia coli O157:H7 Sakai strain. We analysed the RNA polymerase enzyme complex using mass spectrometry and identified associated proteins. Although Escherichia coli O157:H7 Sakai contains more than 1,600 genes not present in the K-12 strain, many of which are predicted to be involved in transcription regulation, all of the identified proteins in this study were coded on the 'core' Escherichia coli genome. A Manuscript describing this work was published: Affinity isolation and I-DIRT mass spectrometric analysis of the Escherichia coli O157:H7 Sakai RNA polymerase complex. Lee DJ, Busby SJ, Westblade LF, Chait BT. J Bacteriol. 2008 Feb;190(4):1284-9.